Dihydroorotate dehydrogenase from Saccharomyces cerevisiae: spectroscopic investigations with the recombinant enzyme throw light on catalytic properties and metabolism of fumarate analogues

In all organisms the fourth catalytic step of the pyrimidine biosynthesis is driven by the flavoenzyme dihydroorotate dehydrogenase (DHODH, EC 1.3.99.11). Cytosolic DHODH of the established model organism Saccharomyces cerevisiae catalyses the oxidation of dihydroorotate to orotate and the reduction of fumarate to succinate. Here, we investigate the structure and mechanism of DHODH from S. cerevisiae and show that the recombinant ScDHODH exists as a homodimeric enzyme in vitro. Inhibition of ScDHODH by the reaction product was observed and kinetic studies disclosed affinity for orotate (K(ic)=7.7 microM; K(ic) is the competitive inhibition constant). The binding constant for orotate was measured through comparison of UV-visible spectra of the bound and unbound recombinant enzyme. The midpoint reduction potential of DHODH-bound flavine mononucleotide determined from analysis of spectral changes was -242 mV (vs. NHE) under anaerobic conditions. A search for alternative electron acceptors revealed that homologues such as mesaconate can be used as electron acceptors.     

微生物燃料电池在毒性物质检测中的应用

微生物燃料电池(Microbial fuel cell,MFC)利用微生物整体作为催化剂催化底物将化学能直接转化为电能,是一种极具应用前景的生物电化学技术。微生物在阳极氧化还原有机物产生电子并传递给阳极,电子通过外电路传递至阴极后将电子释放给阴极中的氧化剂,从而产生电流。当有毒物质进入MFC,微生物活性降低,电子传递量变少,电流降低,而电流的产生与微生物活性呈线性关系,据此可检测样品的毒性。本文主要介绍了微生物燃料电池在毒性物质抗生素、重金属离子、有机污染物、酸等方面的研究,并分析了微生物燃料电池存在的问题及未来研究方向,以期不久的将来微生物燃料电池能付之使用。     

单克隆抗体生产过程中二硫键还原成因及预防方法

单克隆抗体生产过程中二硫键的还原是生物制药领域中的一个常见技术难题,可产生低分子量碎片,影响产品质量,导致蛋白纯度降低、稳定性下降,影响药物的安全性和有效性。抗体二硫键还原实质上是由细胞内的硫氧还蛋白系统和谷胱甘肽系统引起的可逆氧化还原反应,并与具体生产过程参数有关。近年来,随着抗体药物和哺乳动物细胞培养工艺规模的发展,二硫键还原问题频繁发生。为解决此问题,研究人员不断尝试并建立了多种预防方法以保证产品质量。概述了抗体二硫键结构、二硫键还原的主要成因及生产过程中的形成因素,重点阐述了消除或减缓抗体二硫键还原的方法、对策,并列举了几种可行的过程分析技术,以期为单克隆抗体药物生产制造工艺的进一步优化提供参考。     

金属离子胁迫对Sphingomonas sp. Y2降解壬基酚聚氧乙烯醚特性的影响

壬基酚聚氧乙烯醚（NEPOs）是全球应用量最大的非离子型表面活性剂之一,具有环境雌激素毒性。NPEOs的中间代谢产物种类多、难降解,且毒性远高于其母系化合物。为研究金属离子对功能微生物Sphingomonas sp. Y2降解NPEOs特性的影响,分析了金属离子的最低抑制浓度（MIC）、细菌形态、NPEOs降解效率及代谢产物组成等变化。结果显示,菌株Y2对多种金属离子具有耐受性,在重金属培养基中对Mn²⁺、Zn²⁺具有较高的耐受性,MIC分别为500、90 mg/L;在500 mg/L Mn²⁺胁迫下,菌株Y2对NPEOs降解率为100.00%（3 d）;在90 mg/L Zn²⁺胁迫下,菌株Y2对NPEOs的降解率为20.62%（5 d）;两种离子双重胁迫下NPEOs降解率为15.65%（5 d）;Mn²⁺胁迫下菌株Y2细胞表面结构和形态发生明显变化,且改变了NPEOs代谢产物中组分的含量组成,其中短链NPEOs与短链壬基酚聚氧乙烯酸（NPECs）的比例为0.68,与对照相比,抑制/减缓了短链NPEOs的羧化反应。结果表明,菌株Sphingomonas sp. Y2对多种金属离子具有耐受性,Mn²⁺胁迫对细胞表面超微结构及NPEOs中间代谢产物组分组成产生显著影响。该研究将为表面活性剂类污染物的生物降解及相应代谢产物在环境中的毒性评价提供理论依据。     

南方红壤旱坡地花生植株对施用氮肥的响应

选用多年夏花生-冬蔬菜种植制度下的典型红壤旱坡地,通过田间试验,研究了5种氮肥水平下花生根系的形态特征、固氮能力、植株生物量以及荚果产量。结果表明:不同氮肥施用量对花生植株生物量及荚果产量影响均不显著(P=0.091);而不同处理的土壤固氮酶活性与氮肥施用量间呈极显著的负相关(R2=0.88,P=0.005);单株花生根系总长度、总表面积、总分叉数及根尖条数均随氮肥施用量的增加而减小,且与单株花生根瘤生物量呈极显著(P<0.001)正相关。     

短期增温和降水减少对半干旱沙质草地群落特征的影响

【目的】为探究短期增温和降水减少对沙质草地植物群落物种多样性、功能性状及生产力的影响,进一步揭示沙质草地植物群落特征对气候变化的响应。【方法】以科尔沁沙质草地植物群落为研究对象,利用开顶式生长室(OTC)模拟增温,研究降水减少0%、20%、40%和60%与增温的交互作用对沙质草地群落组成、物种多样性和功能性状的影响及群落特征与环境因子关系。【结果】结果显示:（1）短期增温和降水减少使沙质草地植物群落组成和物种优势度发生变化,其中优势物种猪毛蒿和达乌里胡枝子在短期增温和降水减少处理下均明显降低,短期增温和降水减少均显著降低了植物群落盖度和地上生物量。（2）短期增温显著降低了Margalef丰富度指数、Simpson指数、Shannon多样性指数和Pielou均匀度指数,短期降水减少对沙质草地供试4种物种多样性指标均无显著影响。（3）短期增温显著提高了植物群落高度和叶干物质含量,显著降低叶厚度,短期降水减少显著提高了叶干物质含量和叶厚度,显著降低了植株高度和比叶面积。（4）短期增温是影响沙质草地物种多样性和功能性状的主要环境因素。【结论】综上可知,短期增温和降水减少改变了沙质草地群落特征,温度是影响沙质草地群落特征的主要环境因子。     

Multi-Shell Copper Catalysts for Selective Electroreduction of CO2 to Multicarbon Chemicals

Electrocatalytic CO₂ reduction (CO₂R) coupled with renewable electricity has been considered as a promising route for the sustainability transition of energy and chemical industries. However, the unsatisfactory yield of desired products, particularly multicarbon (C₂₊) products, has hindered the implementation of this technology. This work describes a strategy to enhance the yield of C₂₊ product formation in CO₂R by utilizing spatial confinement effects. The finite element simulation results suggest that increasing the number of shells in the catalyst wil lead to a high local concentration of *CO and promotes the formation of C₂₊ products. Inspired by this, Cu nanoparticles are synthesized with desired hollow multi-shell structures. The CO₂ reduction results confirm that as the number of shells increase, the hollow multi-shell copper catalysts exhibit improved selectivity toward C₂₊ products. Specifically, the Cu catalyst with 4.4-shell achieved a high selectivity of over 80% toward C₂₊ at a current density of 900 mA cm⁻². Evidence from in situ attenuated total reflection surface-enhanced infrared absorption spectroscopy unveils that the multi-shell Cu catalyst exhibits an enhanced *CO_atop coverage and the stronger interaction with *CO_atop compared to commercial Cu, confirming the simulation results. Overall, the work promises an effective approach for boosting CO₂R selectivity toward value-added chemicals.     

High-Areal-Capacity and Long-Cycle-Life All-Solid-State Lithium-Metal Battery by Mixed-Conduction Interface Layer

The rapid growth of lithium dendrites has seriously hindered the development and practical application of high-energy-density all-solid-state lithium metal batteries (ASSLMBs). Herein, a soft carbon (SC)-nano Li_6.4La₃Zr_1.4Ta_0.6O₁₂ (LLZTO) (with high ionic conductivity and diffusion coefficient) mixed ionic and electronic conducting interface layer is designed to promote the rapid migration of Li⁺ at the interfacial layer, induce the uniform deposition of lithium metal on nanoscale (nano) LLZTO ion-conducting network inside the interface layer, effectively suppress the growth of lithium dendrites, and significantly improve the electrochemical performance of ASSLMBs. LiZrO₂@LiCoO₂(LZO@LCO)/Li₆PS₅Cl(LPSCl)-nano LLZTO/Li ASSLMB achieves high current density (12.5 mA cm⁻²), ultra-high areal capacity (15 mAh cm⁻², corresponding to LZO@LCO mass loadings of 111.11 mg cm⁻²), and ultra-long cycle life (20 000 cycles). Therefore, the introduction of SC-nano LLZTO mixed conducting interface layer can greatly improve the interfacial stability between solid-state electrolyte (SSE) and lithium metal anode to enable dendrite-free ASSLMBs.     

湘南某矿区蔬菜中Pb、Cd污染状况及健康风险评估

通过采集湘南某矿区周边农田中种植的蔬菜和对应耕作层土壤样品,探究了农田蔬菜Pb、Cd的污染状况,蔬菜和土壤重金属含量之间的相关性,蔬菜中重金属对人体的健康风险。结果表明:(1)污染区土壤重金属污染非常严重,叶菜类蔬菜重金属超出了食品卫生标准限值,但是果菜类蔬菜中重金属没有超出食品卫生标准限值。(2)不同蔬菜不同器官所含Pb、Cd有明显差异;不同种类蔬菜Pb、Cd含量不同,一般是叶菜类果菜类;同种蔬菜不同部位重金属含量的顺序也不同,一般为根茎叶果,或根叶茎果。(3)二类蔬菜中重金属Pb、Cd的富集系数顺序为叶菜类果菜类。(4)蔬菜中Pb、Cd含量与土壤中相对应元素含量均无显著相关性。(5)就农产品安全性而言,果菜类蔬菜比叶菜类蔬菜更适合在此矿区栽培。